RESEARCH OPPORTUNITY AWARD - CHARACTERIZATION OF NAG EXPRESSION AND FUNCTIONS

研究机会奖 - NAG 表达和功能的表征

• 批准号: 8167527
• 负责人: CARLA J GUTHRIDGE
• 金额: $ 1.39万
• 依托单位: UNIVERSITY OF OKLAHOMA HLTH SCIENCES CTR
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-04-01 至 2011-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8167527
• 关键词: Anthrax Vaccines; Anthrax disease; Antibodies; Antigens; Award; Bacillus anthracis; Bacillus anthracis spore; Binding; Biological Assay; Chimeric Proteins; Computer Retrieval of Information on Scientific Projects Database; Environment; Epitopes; Funding; Goals; Grant; Immune response; Immunity; Immunotherapeutic agent; In Vitro; Individual; Institution; Laboratories; Licensing; Maps; Monoclonal Antibodies; Morbidity - disease rate; Peptide Hydrolases; Peptides; Protein Binding; Recombinant Fusion Proteins; Research; Research Personnel; Resources; Serious Adverse Event; Source; Toxin; United States National Institutes of Health; Vaccination; Vaccines; Work; human monoclonal antibodies; improved; in vivo; mortality; three dimensional structure; vaccination strategy

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Bacillus anthracis spores can cause significant morbidity and mortality if released into the environment. There is a need for both an effective vaccine as well as targeted immunotherapeutic agents. The currently licensed vaccine requires six vaccinations, annual boosters, and has been associated with serious adverse events. Thus there has been a major effort made to develop an improved vaccination strategy which can generate lasting protective immunity with reduced vaccinations. Dr. James' laboratory has been working to identify the major humoral targets of B. anthracis that provide protection. Human monoclonal antibodies were previously generated from individuals who received the anthrax vaccine. Each of these monoclonal antibodies has previously been characterized in both in vitro and in vivo toxin neutralization assays as well as by linear epitope and protease fragment mapping studies. Multiple functionally informative anti-PA monoclonal antibodies were identified. However, the structural epitopes recognized by these functionally informative monoclonal antibodies remained unclear. These monoclonal antibodies apparently recognized conformational epitopes rather than the linear peptide epitopes of protective antigen. The goal of this study was to define the protective antigen structural determinants that were specifically recognized by the functionally informative monoclonal anti-PA antibodies. Using the three dimensional structure of anthrax PA as a guide, different domains of PA will be cloned as recombinant fusion proteins. These PA subdomain containing fusion proteins will then be expressed and used in protein binding assays to define which PA domains are required for anti-PA monoclonal antibody binding. The results of these studies will identify which domains of PA are the best targets for generating protective immune responses through vaccination.

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 炭疽芽孢杆菌的孢子如果释放到环境中，会导致严重的发病率和死亡率。既需要有效的疫苗，也需要有针对性的免疫治疗剂。目前获得许可的疫苗需要六次疫苗接种，每年一次，并与严重的不良事件有关。因此，已经作出了重大努力，以制定改进的疫苗接种战略，在减少疫苗接种的情况下产生持久的保护性免疫。詹姆斯博士的实验室一直在努力识别提供保护的炭疽杆菌的主要体液靶标。此前，人类单抗是从接种炭疽疫苗的个人身上产生的。这些单抗都已经在体外和体内毒素中和试验以及线性表位和蛋白酶片段定位研究中得到了表征。鉴定了多种功能信息丰富的抗PA单抗。然而，这些功能信息丰富的单抗识别的结构表位仍然不清楚。这些单抗明显识别构象表位，而不是保护性抗原的线性多肽表位。 这项研究的目的是确定保护性抗原结构决定因素，这些决定因素是由功能信息丰富的抗PA单抗特异性识别的。以炭疽杆菌PA的三维结构为指导，将PA的不同结构域克隆为重组融合蛋白。然后，这些含有PA亚区的融合蛋白将被表达并用于蛋白质结合分析，以确定抗PA单抗结合所需的PA结构域。这些研究的结果将确定PA的哪些区域是通过疫苗接种产生保护性免疫反应的最佳靶点。